Information on EC 3.4.22.49 - separase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY
3.4.22.49
-
RECOMMENDED NAME
GeneOntology No.
separase
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REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
all bonds known to be hydrolysed by this endopeptidase have arginine in P1 and an acidic residue in P4, P6 is often occupied by an acidic residue or by an hydroxy-amino-acid residue, the phosphorylation of which enhances cleavage
show the reaction diagram
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all bonds known to be hydrolysed by this endopeptidase have arginine in P1 and an acidic residue in P4. P6 is often occupied by an acidic residue or by an hydroxy-amino-acid residue, the phosphorylation of which enhances cleavage
show the reaction diagram
unphosphorylatable form of cohesin subunit Rad21 is less efficiently cleaved by Separase
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all bonds known to be hydrolysed by this endopeptidase have arginine in P1 and an acidic residue in P4. P6 is often occupied by an acidic residue or by an hydroxy-amino-acid residue, the phosphorylation of which enhances cleavage
show the reaction diagram
cohesin cleavage by human separase requires DNA in a sequence-nonspecific manner (in vitro). Autocleavage of separase is not stimulated by DNA. Cleavage of the chromosome-associated cohesins is sensitive to nuclease treatment.
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all bonds known to be hydrolysed by this endopeptidase have arginine in P1 and an acidic residue in P4. P6 is often occupied by an acidic residue or by an hydroxy-amino-acid residue, the phosphorylation of which enhances cleavage
show the reaction diagram
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-
-
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REACTION TYPE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
hydrolysis of peptide bond
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-
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hydrolysis of peptide bond
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hydrolysis of peptide bond
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hydrolysis of peptide bond
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hydrolysis of peptide bond
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hydrolysis of peptide bond
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hydrolysis of peptide bond
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SYNONYMS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Cut1/separase
-
-
Esp1
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gene name
Esp1
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Saccharomyces cerevisiae separase homolog, N-terminal region comprises approximately the first 850 amino acids of the 1630 amino acids open reading frame
Esp1
Q03018
included in the Cdc14 early anaphase release network
separase
-
-
separase
Q03018
-
separase
-
-
separase
-
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separin
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-
-
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CAS REGISTRY NUMBER
COMMENTARY
351527-77-0
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ORGANISM
COMMENTARY
LITERATURE
SEQUENCE CODE
SEQUENCE DB
SOURCE
several strains
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Manually annotated by BRENDA team
ecotype Columbia
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Manually annotated by BRENDA team
ESP; gene AtESP
F4JMQ3
UniProt
Manually annotated by BRENDA team
gene Separase
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Manually annotated by BRENDA team
gene Sse/THR, Drosophila separase is encoded by two different genes: (1) Sse-encoding separase-like protein with protease domain and (2) THR (three rows) for the protein interacting with PIM (pimples), a securin homologue of Drosophila
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Manually annotated by BRENDA team
separase is required for sister chromatid separation in mitosis
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Manually annotated by BRENDA team
gene hESP
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Manually annotated by BRENDA team
gene Os02g0770700
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Manually annotated by BRENDA team
gene Separase
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Manually annotated by BRENDA team
commercial baker's yeast
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Manually annotated by BRENDA team
separase is part of the so called Cdc fourteen anaphase release network, that promotes Cdc14 release from the nucleolus during early anaphase
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Manually annotated by BRENDA team
separase mediates Cdc14 release from nucleoli during meiosis through a mechanism that is indepenent of its protease activity
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Manually annotated by BRENDA team
gene XM_002454579
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Manually annotated by BRENDA team
gene LOC100259948
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Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
evolution
F4JMQ3
separases belong to CD clan of cysteine proteases. Unlike other members of this clan, separases are large multidomain proteins with more than 1000 amino acid residues. The catalytic domain of Arabidopsis separase exhibits 31 and 32% identity to the corresponding domains of human and budding yeast homologues, respectively, while the identity exceeds 50% within plant kingdom showing that the proteolytic domain of separases is the most conserved one. The sequence identity drops dramatically for the N-termini of separases. For example, the identity of the first 600 amino acid residues between Arabidopsis and Vitis vinifera separases does not exceed 39%, and it is only 30% between Arabidopsis and rice. Mode of action in vivo and mechanistic differences in mitosis between organisms, overview
evolution
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separases belong to CD clan of cysteine proteases. Unlike other members of this clan, separases are large multidomain proteins with more than 1000 amino acid residues. Mode of action in vivo and mechanistic differences in mitosis between organisms, overview
evolution
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separases belong to CD clan of cysteine proteases. Unlike other members of this clan, separases are large multidomain proteins with more than 1000 amino acid residues. The catalytic domain of Arabidopsis thaliana separase exhibits 31 and 32% identity to the corresponding domains of human and budding yeast homologues, respectively. The sequence identity drops dramatically for the N-termini of separases. Mode of action in vivo and mechanistic differences in mitosis between organisms, overview
evolution
-
separases belong to CD clan of cysteine proteases. Unlike other members of this clan, separases are large multidomain proteins with more than 1000 amino acid residues. The sequence identity exceeds 50% within plant kingdom showing that the proteolytic domain of separases is the most conserved one. The sequence identity drops dramatically for the N-termini of separases. For example, the identity of the first 600 amino acid residues between Arabidopsis thaliana and Oryza sativa is only 30%. Mode of action in vivo and mechanistic differences in mitosis between organisms, overview
evolution
-
separases belong to CD clan of cysteine proteases. Unlike other members of this clan, separases are large multidomain proteins with more than 1000 amino acid residues. Mode of action in vivo and mechanistic differences in mitosis between organisms, overview
evolution
-
separases belong to CD clan of cysteine proteases. Unlike other members of this clan, separases are large multidomain proteins with more than 1000 amino acid residues. The catalytic domain of Arabidopsis thaliana separase exhibits 31 and 32% identity to the corresponding domains of human and budding yeast homologues, respectively. The sequence identity drops dramatically for the N-termini of separases. Mode of action in vivo and mechanistic differences in mitosis between organisms, overview
evolution
-
separases belong to CD clan of cysteine proteases. Unlike other members of this clan, separases are large multidomain proteins with more than 1000 amino acid residues. Mode of action in vivo and mechanistic differences in mitosis between organisms, overview
evolution
-
separases belong to CD clan of cysteine proteases. Unlike other members of this clan, separases are large multidomain proteins with more than 1000 amino acid residues. The sequence identity exceeds 50% within plant kingdom showing that the proteolytic domain of separases is the most conserved one. The sequence identity drops dramatically for the N-termini of separases. For example, the identity of the first 600 amino acid residues between Arabidopsis thaliana and Vitis vinifera separases does not exceed 39%. Mode of action in vivo and mechanistic differences in mitosis between organisms, overview
malfunction
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loss of separase function during the early mitotic divisions causes cytokinesis failure, depletion of separase causes the accumulation of RAB-11-positive vesicles at the cleavage furrow and midbody
malfunction
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loss of separase blocks centriole disengagement during mitotic exit and delays assembly of new centrioles during the following S phase
malfunction
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Arabidopsis thaliana radially swollen 4 (rsw4), a temperature-sensitive mutant, harbors a mutation in At4g22970, the separase. Loss of separase function in rsw4 at the restrictive temperature is indicated by the widespread failure of replicated chromosomes to disjoin. rsw4 has neither pronounced cell cycle arrest nor anomalous spindle formation, rsw4 roots have disorganized cortical microtubules and accumulate the mitosis-specific cyclin, cyclin B1,1
malfunction
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meiotic expression of ESP RNA interference blocks the removal of cohesin during both meiosis I and II, results in alterations in nonhomologous centromere association, disrupts the radial microtubule system after telophase II, and affects the proper establishment of nuclear cytoplasmic domains, resulting in the formation of multinucleate microspores
malfunction
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cells that do not express both Cdc55 and securin prematurely separate their sister chromatids, leading to cell death
malfunction
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cells that do not express both Cdc55 and securin prematurely separate their sister chromatids, leading to cell death. Mutant mice lacking securin and expressing a non-phosphorylatable separase die in embryonic stage. But mouse embryonic stem cells lacking both these separase regulations can still progress through mitosis in a timely fashion with correct chromosome segregation
malfunction
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cells that do not express both Cdc55 and securin prematurely separate their sister chromatids, leading to cell death
malfunction
F4JMQ3
knocking down AtESP in meiocytes using RNAi unexpectedly converts the symmetric radial microtubule systems that form after telophase II into asymmetric structures partially resembling phragmoplasts
malfunction
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loss of either APC or separase results in a failure of the transduction of the presumed polarity signal from the centrosome cortex
malfunction
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human cells with one hESP allele-encoding uncleavable protein and another allele harboring a single cleavage site grow slowly owing to cell cycle delay, in particular during G2/M transition, but not when it was expected, i.e. during anaphase
malfunction
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in the cells lacking securin Pds1, Esp1 distribution is largely restricted to the cytoplasm
malfunction
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cells depleted of securin or separase display defective acidification of early endosomes and increased membrane recruitment of vacuolar ATPase complexes, mimicking the effect of the specific V-ATPase inhibitor Bafilomycin A1. Securin and separase depletion causes trans-Golgi network and endosome swelling independent of cell cycle. Endosome-mediated receptor degradation and recycling are also significantly impaired by securin and separase depletion, although not receptor internalization or Rab5 activity and autophagy
physiological function
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separase plays a pivotal role in the separation of sister chromatids at anaphase by cleaving its substrate cohesin Rad21
physiological function
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separase is required for cytokinesis by regulating the incorporation of RAB-11-positive vesicles into the plasma membrane at the cleavage furrow and midbody
physiological function
-
separase acts during M phase to license centrosome duplication
physiological function
-
plant separase, in addition to cleaving cohesin, regulates cyclin B1,1, with profound ramifications for morphogenesis
physiological function
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sister chromatid separation at anaphase is triggered by cleavage of the cohesin subunit Scc1, which is mediated by separase
physiological function
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Separase is essential for sister chromatid separation during anaphase II. Separase-mediated proteolytic cleavage of the alpha-kleisin subunit of the cohesin complex at the metaphase-to-anaphase transition is essential for the proper segregation of chromosomes. Separase is also involved in mitotic and meiotic anaphase spindle assembly and elongation, interphase spindle pole body positioning, and epithelial cell reorganization
physiological function
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separase Esp1 is a protease specialized in the cleavage of sister chromatid cohesion. When inhibitor securin Pds1 is degraded, Esp1 is activated, and cells transit into anaphase. Esp1, together with Clb2- and Polo-kinases, promotes Cdc14 activation through the FEAR network. Separase also leads to the activation of Cdc14 phosphatase. The phosphatase is kept inactive in the nucleolus by Net1 throughout the cell cycle until anaphase. The proteolytic function of separase causes spindle elongation by cohesin cleavage, which activates mitotic exit network, MEN, by bringing Tem1 together with its activator Lte1
physiological function
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sister chromatid cohesion depends on the cohesin complex, a proteinaceous ring that entraps the chromatids together. At the metaphase-to-anaphase transition, separase is activated and completely dissolves the cohesion by cleaving SCC1, a subunit of the cohesin complex. As one of the key executors of anaphase, separase is regulated temporally and spatially by often redundant mechanisms. Chromosomal DNA dependent cohesin cleavage by separase is a component of a regulatory pathway that cells utilize to protect the bulk of cohesin. Degradation of securin plays a critical role in the timely activation of separase activity. In vertebrate cells, separase is phosphorylated and inhibited before anaphase by a cyclin B/CDK1. Nuclear exclusion of separase might provide the means to preclude cohesin cleavage at telophase and G1 stage of the cell cycle
physiological function
-
sister chromatid cohesion depends on the cohesin complex, a proteinaceous ring that entraps the chromatids together. At the metaphase-to-anaphase transition, separase is activated and completely dissolves the cohesion by cleaving SCC1, a subunit of the cohesin complex. As one of the key executors of anaphase, separase is regulated temporally and spatially by often redundant mechanisms. Chromosomal DNA dependent cohesin cleavage by separase is a component of a regulatory pathway that cells utilize to protect the bulk of cohesin. Degradation of securin plays a critical role in the timely activation of separase activity. But securin-independent separase regulation occur, cohesin cleavage is inhibited by a PP2ACdc55-dependent mechanism. Most of the budding yeast cohesin is cleaved in anaphase, and this cleavage is stimulated by phosphorylation of the Scc1 subunit by the Plk1 kinase
physiological function
F4JMQ3
function of separases in metaphase to anaphase transition, overview. Separase cleaves and removes the remaining centromeric cohesin. In plants, the molecular mechanisms regulating sister chromatid separation remain largely elusive. AtESP plays a role in microtubule organization or cell polarity, and an additional role for AtESP beyond cohesin cleavage
physiological function
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function of separases in metaphase to anaphase transition, overview
physiological function
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function of separases in metaphase to anaphase transition, overview. The activated APCCdc20/separase pathway plays a fundamental role in the establishment of the anterior-posterior axis
physiological function
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function of separases in metaphase to anaphase transition, overview. Human separase is a potential oncogene and hESP transcripts are accumulated in a large number of tumors
physiological function
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function of separases in metaphase to anaphase transition, overview. Separase cleaves and removes the remaining centromeric cohesin. In plants, the molecular mechanisms regulating sister chromatid separation remain largely elusive
physiological function
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function of separases in metaphase to anaphase transition, overview. Separase cleaves and removes the remaining centromeric cohesin. In yeasts, separase is responsible for the removal of both arm and centromeric cohesin after its phosphorylation by Cdc5 or other Plks. Esp1 action is not limited to this stage. When securin is depleted in yeast cells, the proteolytic activity of Esp1 is no longer cell cycle regulated, while Scc1 is cleaved on schedule suggesting the existence of additional regulatory elements
physiological function
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function of separases in metaphase to anaphase transition, overview. Separase cleaves and removes the remaining centromeric cohesin. In yeasts, separase is responsible for the removal of both arm and centromeric cohesin after its phosphorylation by Cdc5 or other Plks. Separase can target both centromeric cohesin and cohesin of chromosomal arms. Cohesin is implicated in transcriptional regulation in Schizosaccharomyces pombe. When securin is depleted in yeast cells, the proteolytic activity of Esp1 is no longer cell cycle regulated, while Scc1 is cleaved on schedule suggesting the existence of additional regulatory elements
physiological function
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function of separases in metaphase to anaphase transition, overview. Separase cleaves and removes the remaining centromeric cohesin. In plants, the molecular mechanisms regulating sister chromatid separation remain largely elusive
physiological function
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chromosomal segregation is mediated by cyclin-dependent kinase 1 and separase, which is regulated by cell division cycle 6, Cdc6, a mitotic substrate of polo-like kinase 1. The phosphorylation of Cdc6 by Plk1 regulates the activity of separase through the association with Cdk1
physiological function
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functional role of securin and separase in the modulation of membrane traffic and protein secretion implicating regulation of V-ATPase assembly and function. Separase activity is controlled by securin, i.e. pituitary tumor transforming gene 1, PTTG1, a member of a divergent class of anaphase inhibitors whose proteosomal degradation by the anaphase promoting complex, APC, is required to release separase and allow its activation
metabolism
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Plk1-mediated phosphorylation of Cdc6 on residue T37 promotes the interaction of Cdc6 and Cdk1, leading to the attenuation of Cdk1 activity, release of separase, and subsequent anaphase progression
additional information
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dynamics of the mitotic exit control system in budding yeast, Queralt's model, modifications, overview. Queralt's model centres around the non-proteolytic function of separase Esp1, which triggers a positive feedback in the activation of MEN by FEAR-induced release of Cdc14
additional information
F4JMQ3
securin, in addition to its inhibitory role, can act as a molecular chaperone of separase, essential for its proper folding
additional information
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securin, in addition to its inhibitory role, can act as a molecular chaperone of separase, essential for its proper folding
additional information
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human separase is present in cells as a part of very large protein complex, which in addition to securin contains also Cdk and cyclin B1, both able to inhibit separase. Securin, in addition to its inhibitory role, can act as a molecular chaperone of separase, essential for its proper folding. The human separase-securin complex shows a whale-type distinct elongated pattern. In this complex, securin is thought to interact with the N-part of separase spanned by the ARM repeats. The N- to C-terminus intramolecular interaction in separase molecules is considered to be necessary for their catalytic activation, and this interaction is abolished by securin binding
additional information
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securin, in addition to its inhibitory role, can act as a molecular chaperone of separase, essential for its proper folding
additional information
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securin, in addition to its inhibitory role, can act as a molecular chaperone of separase, essential for its proper folding. Securin is dispensable for the growth of normal human cells. The first 156 amino acids of Esp1 seem imperative for the binding of securin Pds1, it interacts with other parts of Esp1 as well. Securin is not only a guardian of separase, but is also responsible for its translocation to the nucleus in the budding yeast
additional information
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securin, in addition to its inhibitory role, can act as a molecular chaperone of separase, essential for its proper folding. Interaction takes place between the N-terminus of separase and the C-terminus of securin
additional information
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securin, in addition to its inhibitory role, can act as a molecular chaperone of separase, essential for its proper folding
additional information
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cells expressing wild-type Cdc6 display lower Cdk1 activity and higher separase activity than cells expressing Cdc6 mutant T37V
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
acetyl-Asp-Arg-Glu-Ile-Nle-Arg-7-amido-4-methylcoumarin + H2O
acetyl-Asp-Arg-Glu-Ile-Nle-Arg + 7-amino-4-methylcoumarin
show the reaction diagram
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acetyl-Asp-Arg-Glu-Ile-Nle-Arg is the Rad21-peptidyl
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?
Cdc14 + H2O
?
show the reaction diagram
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separase regulates INCENP-Aurora B anaphase spindle function through activation of Cdc14
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-
?
cohesin + H2O
fragments of cohesin
show the reaction diagram
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-
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cohesin + H2O
fragments of cohesin
show the reaction diagram
-
-
-
?
cohesin + H2O
fragments of cohesin
show the reaction diagram
-
-
-
-
?
cohesin + H2O
fragments of cohesin
show the reaction diagram
-
-
-
-
cohesin + H2O
fragments of cohesin
show the reaction diagram
-
-
-
-
cohesin + H2O
fragments of cohesin
show the reaction diagram
-
-
-
-
?
cohesin + H2O
fragments of cohesin
show the reaction diagram
-
-
-
-
?
cohesin + H2O
fragments of cohesin
show the reaction diagram
-
-
-
-
cohesin + H2O
fragments of cohesin
show the reaction diagram
-
-
-
?
cohesin + H2O
fragments of cohesin
show the reaction diagram
-
separase Esp1 is implicated in the removal of cohesin which links sister chromatids in the S-phase and is postulated to be a specific protease or a positive regulator of protease against cohesin
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?
cohesin + H2O
fragments of cohesin
show the reaction diagram
-
-
-
-
?
cohesin + H2O
cleaved cohesin
show the reaction diagram
-
-
-
-
-
cohesin + H2O
cleaved cohesin
show the reaction diagram
-
-
-
-
-
cohesin + H2O
cleaved cohesin
show the reaction diagram
-
-
-
-
?
cohesin + H2O
cleaved cohesin
show the reaction diagram
-
-
-
-
-
cohesin + H2O
cleaved cohesin
show the reaction diagram
-
separase is required for sister chromatid separation during mitosis
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-
-
cohesin + H2O
cleaved cohesin
show the reaction diagram
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separase, i.e. Esp1, is implicated in the removal of cohesin which links sister chromatids in the S-phase and is postulated to be a specific protease or a positive regulator of protease against cohesin
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-
cohesin + H2O
cleaved cohesin
show the reaction diagram
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separase triggers anaphase by cohesin cleavage, and triggers the release and activation of the phosphatase Cdc14 independently of its protease activity
-
-
-
cohesin + H2O
?
show the reaction diagram
-
-
-
-
?
cohesin + H2O
?
show the reaction diagram
-
-
-
-
?
cohesin + H2O
?
show the reaction diagram
-
-
-
-
?
cohesin + H2O
?
show the reaction diagram
F4JMQ3
-
-
-
?
cohesin + H2O
?
show the reaction diagram
-
-
-
-
?
cohesin + H2O
?
show the reaction diagram
-
separase-mediated cleavage of cohesin at interphase is required for DNA repair. It is proposed that the securin-separase complex might aid DNA repair by removing local cohesin in interphase cells
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-
?
cohesin + H2O
?
show the reaction diagram
-
cohesin cleavage by human separase requires DNA in a sequence-nonspecific manner
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-
?
cohesin + H2O
?
show the reaction diagram
-
separase selectively cleaves only the chromosome-associated cohesin with the cofactor chromosomal DNA
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-
?
Rad 21 + H2O
?
show the reaction diagram
-
-
-
-
?
Rad21 + H2O
fragments of Rad21
show the reaction diagram
-
-
-
-
?
Rad21 + H2O
fragments of Rad21
show the reaction diagram
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Rad21 is cleaved by the separase Cut1 at the metaphase-anaphase transition
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?
Rad21 + H2O
cleaved Rad21
show the reaction diagram
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Rad21 is cleaved by Cut1 at the metaphase-anaphase transition
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-
Rec8 + H2O
fragments of Rac8
show the reaction diagram
-
-
-
?
Rec8 + H2O
fragments of Rac8
show the reaction diagram
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cleavage of meiosis-specific Scc1 variant Rec8 is required for chromosome segregation during meiosis
-
?
Rec8 + H2O
cleaved Rac8
show the reaction diagram
-
-
-
-
-
Rec8 + H2O
cleaved Rac8
show the reaction diagram
-
cleavage of meiosis-specific Scc1 variant Rec8 is required for chromosome segregation during meiosis
-
-
-
Rec8 + H2O
?
show the reaction diagram
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-, cleavage is required for meiotic nuclear divisions in fission yeast
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-
?
Rec8 + H2O
?
show the reaction diagram
P60330
resolution of chiasmata in oocytes requires separase-mediated proteolysis. Proteolytic cleavage by separase is essential for Rec8's removal from chromosome arms and for chiasma resolution but not for the first polar body
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-
?
Rec8 + H2O
fragments of Rec8
show the reaction diagram
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phosphorylation either of separase or cohesin is necessary for Rec8 cleavage
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-
-
Rec8 + H2O
fragments of Rec8
show the reaction diagram
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the phosphorylations of Rec8 at S412 and the surrounding sites, which are most probably dependent on CK1, are essential for the cleavability of Rec8 by separase
-
-
?
Rec8 + H2O
fragments of Rec8
show the reaction diagram
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Rec8 is a meiosis-specific alpha-kleisin subunit of cohesion, phosphorylation by polo-like kinase 1 of either Rec8 or separase, or both, promotes Rec8 cleavage. Separase prefers to cleave at R454
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-
?
Rec8 + H2O
fragments of Rec8
show the reaction diagram
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phosphorylation either of separase or cohesin is necessary for Rec8 cleavage
-
-
-
Scc1 + H2O
fragments of Scc1
show the reaction diagram
-
-
-
-
Scc1 + H2O
fragments of Scc1
show the reaction diagram
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separase Esp1 cleaves Scc1, the central subunit of the chromosomal cohesin complex during mitosis
-
?
Scc1 + H2O
fragments of Scc1
show the reaction diagram
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Scc1 is a subunit of cohesion, centrosomal Scc1 is cleaved by separase coincidentally with chromatin Scc1
-
-
?
Scc1 + H2O
fragments of Scc1
show the reaction diagram
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Scc1 is an alpha-kleisin subunit of cohesin
-
-
?
Scc1 + H2O
cleaved Scc1
show the reaction diagram
-
-
-
-
-
Scc1 + H2O
cleaved Scc1
show the reaction diagram
-
-
-
-
-
Scc1 + H2O
cleaved Scc1
show the reaction diagram
-
-
-
-
-
Scc1 + H2O
cleaved Scc1
show the reaction diagram
-
Esp1 cleaves Scc1, the central subunit of the chromosomal cohesin complex during mitosis
-
-
-
Scc1 + H2O
cleaved Scc1
show the reaction diagram
-
Scc1 contains 2 separase cleavage sites
-
-
-
Scc1 + H2O
cleaved Scc1
show the reaction diagram
-
cleavage at 2 defined sites results in rapid destabilization of Scc1 and its dissociation from chromosomes
-
-
-
Slk19 + H2O
fragments of Slk19
show the reaction diagram
-
separase cleaves the kinetochore-associated protein Slk19 in anaphase
-
?
Slk19 + H2O
cleaved Slk19
show the reaction diagram
-
-
-
-
-
Slk19 + H2O
cleaved Slk19
show the reaction diagram
-
separase cleaves the kinetochore-associated protein Slk19 in metaphase-to-anaphase transition
-
-
-
Slk19 + H2O
?
show the reaction diagram
-
a protein implicated in the mitotic exit via its role in the stabilization of spindle in budding yeast
-
-
?
SYN1 + H2O
?
show the reaction diagram
-, Q5IBC5
the enzyme plays an essential role in embryo development. The enzyme is required for the removal of cohesin from meiotic chromosomes. The cleavage of SYN1 by separase is responsible for the release of sister chromatid cohesion during meiosis
-
-
?
cohesin + H2O
?
show the reaction diagram
-
separase selectively cleaves only the chromosome-associated cohesin with the cofactor chromosomal DNA, chromosome-associated cohesin is cleaved preferentially by separase, mechanism
-
-
?
additional information
?
-
-
auto-cleavage of separase coordinates multiple aspects of the G2/M programme in human cells, thus contributing to the timing and efficiency of chromosome segregation
-
-
-
additional information
?
-
-
deletion of separase specifically blocks sister chromatid separation but not other aspects of mitosis, mitotic exit, cytokinesis, or even chromosome replication
-
-
-
additional information
?
-
-
downregulation of PP2A(Cdc55) phosphatase by separase initiates mitotic exit. At anaphase onset, PP2A(Cdc55) activity is downregulated in a separase-dependent manner, triggering a first wave of Cdk-dependent Net1 phosphorylation and Cdc14 release. Separase is the essential trigger for Cdc14 activation in anaphase
-
-
-
additional information
?
-
-
Hela cells lacking separase are delayed or arrested at the G2-M phase transition. Without separase, cells also have a prolonged prometaphase. Separase is required at multiple cell cycle stages including the interphase. Its role in promoting loss of sister chromatid cohesion might be important preferentially at arms but not centromers
-
-
-
additional information
?
-
-
MAP kinase is required for the increased securin-separase interaction that rescues separase mutants under stresses
-
-
-
additional information
?
-
-
separase auto-cleavage coordinates multiple aspects of the G2/M programme in human cells, thus contributing to the timing and efficiency of chromosome segregation
-
-
-
additional information
?
-
-
separase is a stoichiometric inhibitor of cyclin-dependent kinase 1. This function of separase is independent of proteolytic activity
-
-
-
additional information
?
-
-
separase is primarily required for sister chromatid separation during mitotic and meiotic division, by cleavage of a subunit of the cohesin complex. Separase is also and perhaps indirectly important for epithelial integrity
-
-
-
additional information
?
-
-
separase shows self-cleavage upon activation. Separase is auto-cleaved between meiosis I and II in oocytes
-
-
-
additional information
?
-
-
the meiosis I-to-meiosis II transition in mouse oocytes requires separase activity
-
-
-
additional information
?
-
-
Cdc48 is required for the stability of Cut1/separase in mitotic anaphase
-
-
-
additional information
?
-
-
separase recognizes both, a cleavage site consensus sequence as well as features outside the cleavage site
-
-
-
additional information
?
-
-
TbSep is essential for the segregation of both large and minichromosomes (RNAi experiment)
-
-
-
additional information
?
-
-
does not cleave Leu-Glu-His-Asp-CHO-7-amido-4-methylcoumarin
-
-
-
additional information
?
-
-
autocleavage of human separase is to be essential and results in conformational changes
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
Cdc14 + H2O
?
show the reaction diagram
-
separase regulates INCENP-Aurora B anaphase spindle function through activation of Cdc14
-
-
?
cohesin + H2O
fragments of cohesin
show the reaction diagram
-
-
-
-
?
cohesin + H2O
fragments of cohesin
show the reaction diagram
-
-
-
-
?
cohesin + H2O
cleaved cohesin
show the reaction diagram
-
-
-
-
-
cohesin + H2O
cleaved cohesin
show the reaction diagram
-
-
-
-
-
cohesin + H2O
cleaved cohesin
show the reaction diagram
-
-
-
-
?
cohesin + H2O
cleaved cohesin
show the reaction diagram
-
-
-
-
-
cohesin + H2O
cleaved cohesin
show the reaction diagram
-
separase is required for sister chromatid separation during mitosis
-
-
-
cohesin + H2O
cleaved cohesin
show the reaction diagram
-
separase, i.e. Esp1, is implicated in the removal of cohesin which links sister chromatids in the S-phase and is postulated to be a specific protease or a positive regulator of protease against cohesin
-
-
-
cohesin + H2O
cleaved cohesin
show the reaction diagram
-
separase triggers anaphase by cohesin cleavage, and triggers the release and activation of the phosphatase Cdc14 independently of its protease activity
-
-
-
cohesin + H2O
?
show the reaction diagram
-
-
-
-
?
cohesin + H2O
?
show the reaction diagram
F4JMQ3
-
-
-
?
cohesin + H2O
?
show the reaction diagram
-
-
-
-
?
cohesin + H2O
?
show the reaction diagram
-
separase-mediated cleavage of cohesin at interphase is required for DNA repair. It is proposed that the securin-separase complex might aid DNA repair by removing local cohesin in interphase cells
-
-
?
cohesin + H2O
?
show the reaction diagram
-
cohesin cleavage by human separase requires DNA in a sequence-nonspecific manner
-
-
?
cohesin + H2O
?
show the reaction diagram
-
separase selectively cleaves only the chromosome-associated cohesin with the cofactor chromosomal DNA
-
-
?
cohesin + H2O
?
show the reaction diagram
-
separase selectively cleaves only the chromosome-associated cohesin with the cofactor chromosomal DNA, chromosome-associated cohesin is cleaved preferentially by separase, mechanism
-
-
?
Rad 21 + H2O
?
show the reaction diagram
-
-
-
-
?
Rad21 + H2O
fragments of Rad21
show the reaction diagram
-
-
-
-
?
Rad21 + H2O
cleaved Rad21
show the reaction diagram
-
Rad21 is cleaved by Cut1 at the metaphase-anaphase transition
-
-
-
Rec8 + H2O
cleaved Rac8
show the reaction diagram
-
-
-
-
-
Rec8 + H2O
cleaved Rac8
show the reaction diagram
-
cleavage of meiosis-specific Scc1 variant Rec8 is required for chromosome segregation during meiosis
-
-
-
Rec8 + H2O
?
show the reaction diagram
-
cleavage is required for meiotic nuclear divisions in fission yeast
-
-
?
Rec8 + H2O
?
show the reaction diagram
P60330
resolution of chiasmata in oocytes requires separase-mediated proteolysis. Proteolytic cleavage by separase is essential for Rec8's removal from chromosome arms and for chiasma resolution but not for the first polar body
-
-
?
Rec8 + H2O
fragments of Rec8
show the reaction diagram
-
phosphorylation either of separase or cohesin is necessary for Rec8 cleavage
-
-
-
Rec8 + H2O
fragments of Rec8
show the reaction diagram
-
the phosphorylations of Rec8 at S412 and the surrounding sites, which are most probably dependent on CK1, are essential for the cleavability of Rec8 by separase
-
-
?
Rec8 + H2O
fragments of Rec8
show the reaction diagram
-
phosphorylation either of separase or cohesin is necessary for Rec8 cleavage
-
-
-
Scc1 + H2O
fragments of Scc1
show the reaction diagram
-
Scc1 is a subunit of cohesion, centrosomal Scc1 is cleaved by separase coincidentally with chromatin Scc1
-
-
?
Scc1 + H2O
cleaved Scc1
show the reaction diagram
-
-
-
-
-
Scc1 + H2O
cleaved Scc1
show the reaction diagram
-
-
-
-
-
Scc1 + H2O
cleaved Scc1
show the reaction diagram
-
-
-
-
-
Scc1 + H2O
cleaved Scc1
show the reaction diagram
-
Esp1 cleaves Scc1, the central subunit of the chromosomal cohesin complex during mitosis
-
-
-
Scc1 + H2O
cleaved Scc1
show the reaction diagram
-
Scc1 contains 2 separase cleavage sites
-
-
-
Scc1 + H2O
cleaved Scc1
show the reaction diagram
-
cleavage at 2 defined sites results in rapid destabilization of Scc1 and its dissociation from chromosomes
-
-
-
Slk19 + H2O
cleaved Slk19
show the reaction diagram
-
-
-
-
-
Slk19 + H2O
cleaved Slk19
show the reaction diagram
-
separase cleaves the kinetochore-associated protein Slk19 in metaphase-to-anaphase transition
-
-
-
Slk19 + H2O
?
show the reaction diagram
-
a protein implicated in the mitotic exit via its role in the stabilization of spindle in budding yeast
-
-
?
SYN1 + H2O
?
show the reaction diagram
-, Q5IBC5
the enzyme plays an essential role in embryo development. The enzyme is required for the removal of cohesin from meiotic chromosomes. The cleavage of SYN1 by separase is responsible for the release of sister chromatid cohesion during meiosis
-
-
?
cohesin + H2O
fragments of cohesin
show the reaction diagram
-
-
-
-
?
additional information
?
-
-
auto-cleavage of separase coordinates multiple aspects of the G2/M programme in human cells, thus contributing to the timing and efficiency of chromosome segregation
-
-
-
additional information
?
-
-
deletion of separase specifically blocks sister chromatid separation but not other aspects of mitosis, mitotic exit, cytokinesis, or even chromosome replication
-
-
-
additional information
?
-
-
downregulation of PP2A(Cdc55) phosphatase by separase initiates mitotic exit. At anaphase onset, PP2A(Cdc55) activity is downregulated in a separase-dependent manner, triggering a first wave of Cdk-dependent Net1 phosphorylation and Cdc14 release. Separase is the essential trigger for Cdc14 activation in anaphase
-
-
-
additional information
?
-
-
Hela cells lacking separase are delayed or arrested at the G2-M phase transition. Without separase, cells also have a prolonged prometaphase. Separase is required at multiple cell cycle stages including the interphase. Its role in promoting loss of sister chromatid cohesion might be important preferentially at arms but not centromers
-
-
-
additional information
?
-
-
MAP kinase is required for the increased securin-separase interaction that rescues separase mutants under stresses
-
-
-
additional information
?
-
-
separase auto-cleavage coordinates multiple aspects of the G2/M programme in human cells, thus contributing to the timing and efficiency of chromosome segregation
-
-
-
additional information
?
-
-
separase is a stoichiometric inhibitor of cyclin-dependent kinase 1. This function of separase is independent of proteolytic activity
-
-
-
additional information
?
-
-
separase is primarily required for sister chromatid separation during mitotic and meiotic division, by cleavage of a subunit of the cohesin complex. Separase is also and perhaps indirectly important for epithelial integrity
-
-
-
additional information
?
-
-
separase shows self-cleavage upon activation. Separase is auto-cleaved between meiosis I and II in oocytes
-
-
-
additional information
?
-
-
the meiosis I-to-meiosis II transition in mouse oocytes requires separase activity
-
-
-
additional information
?
-
-
TbSep is essential for the segregation of both large and minichromosomes (RNAi experiment)
-
-
-
additional information
?
-
-
autocleavage of human separase is to be essential and results in conformational changes
-
-
-
METALS and IONS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
Ca2+
-
Ca2+-levels affect separase function, C-terminal region contains a Ca2+-binding motif
Ca2+
F4JMQ3
the enzyme contains a Ca2+-binding EF-hand motif, which can possibly affect separase interaction with the spindle, similar to the budding yeast Esp1, or alternatively Ca2+ might be a critical component for (auto-)catalysis
INHIBITORS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
Cdc55
-
results suggest that Cdc55 acts as inhibitor downstream from shugoshin
-
Pds1
Q03018
Pds1 is a chaperone, inhibition of Esp1 by overexpression of undegradable Pds1 blocks mitotic exit via blockage of cohesin cleavage
-
peptide
-
acyloxymethyl ketone derivative of human SCC1 cleavage site peptide, chloromethyl ketone derivatives of the yeast Scc1 cleavage site
securin
-
securin homolog pimples, i.e. PIM, binds and inhibits separase
securin
-
small protein that binds to and inhibits separase until all pairs of chromatids have established bipolar spindle attachments
securin
-
inhibits the proteolytic activity of separase
securin
-
securin inhibits separase by blocking the access of substrates to the active site
securin
-
securin regulates both the proteolytic and non-proteolytic activities of separase
securin
-
destruction of securin occurs only upon the correct attachment of chromosomes to the spindle
securin
-
activity inhibited prior to the onset of the anaphase
securin
F4JMQ3
in addition to its inhibitory role, can act as a molecular chaperone of separase, essential for its proper folding
securin
-
in addition to its inhibitory role, can act as a molecular chaperone of separase, essential for its proper folding
securin
-
in addition to its inhibitory role, can act as a molecular chaperone of separase, essential for its proper folding. Securin is dispensable for the growth of normal human cells, in contrast to cancer cells, where depletion of PTTG1 leads to chromosome instability. The human separase-securin complex shows a whale-type distinct elongated pattern. In this complex, securin is thought to interact with the N-part of separase spanned by the ARM repeats. The N- to C-terminus intramolecular interaction in separase molecules is considered to be necessary for their catalytic activation, and this interaction is abolished by securin binding
securin
-
in addition to its inhibitory role, can act as a molecular chaperone of separase, essential for its proper folding
securin
-
in addition to its inhibitory role, can act as a molecular chaperone of separase, essential for its proper folding. The first 156 amino acids of Esp1 seem imperative for the binding of securin Pds1, it interacts with other parts of Esp1 as well
securin
-
in addition to its inhibitory role, can act as a molecular chaperone of separase, essential for its proper folding. Interaction takes place between the N-terminus of separase and the C-terminus of securin
securin
-
in addition to its inhibitory role, can act as a molecular chaperone of separase, essential for its proper folding
securin Pds1
-
an inhibitor of separase Esp1 in budding yeast. As Pds1 is degraded, Esp1 is activated, and cells transit into anaphase
-
shugoshin
-
prevents separase activation independently of securin, protein phosphatase 2A coupled to regulatory subunit Cdc55 is essential for Shugoshin-mediated inhibition
-
Cdk1 kinase
-
phosphorylation-dependent binding of the cyclin B1 subunit of Cdk1 kinase to a Cdc6-like domain of separase inhibits separase
-
additional information
-
nuclear exclusion of separase might provide the means to preclude cohesin cleavage at telophase and G1 stage of the cell cycle
-
additional information
-
cohesin cleavage is inhibited by a PP2ACdc55-dependent mechanism
-
additional information
-
separase is kept inactive in human cells by Cdk(Cdc2)-dependent phosphorylation even when securin is degraded
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
cell division cycle 6
-
Cdc6, a mitotic substrate of polo-like kinase. Cleavage of chesin/Rad21 is much greater in wild-type Cdc6 expressing cells than in GFP and Cdc6-T37V mutant expressing cells
-
DNA
-
chromosomal DNA is required as a cofactor for the cleavage of cohesin to occur, and allows separase to selectively cleave only the chromosome-associated cohesin. Separase binds to DNA in a sequence nonspecific manner in vitro and associates with the entire length of the mitotic chromosomes
securin
-
activation may be due to separase localization
securin
-
securin is required to support separase activity in anaphase
securin
-
the central domain of securin has a functionally essential specific sequence that may directly interact with the catalytic region of separase. This central securin domain is unrelated to destruction by polyubiquitination, but essential for the activation of separase
DNA
-
chromosomal DNA is required as a cofactor for the cleavage of cohesin to occur, and allows separase to selectively cleave only the chromosome-associated cohesin
additional information
-
separase shows self-cleavage upon activation
-
additional information
-
separase can be activated by incubating immunoglobulin G-Sepharose 6 containing ZZ TEV4-separase with anaphase-initiated Xenopus cytostatic factor to degrade its inhibitor securin at room temperature for 1 h
-
additional information
-
Cdc20 is essential for mitotic progression
-
additional information
-
most of the budding yeast cohesin is cleaved in anaphase, and this cleavage is stimulated by phosphorylation of the Scc1 subunit by the Plk1 kinase
-
KM VALUE [mM]
KM VALUE [mM] Maximum
SUBSTRATE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
IMAGE
0.293
-
acetyl-Asp-Arg-Glu-Ile-Nle-Arg-7-amido-4-methylcoumarin
-
at 37C
SPECIFIC ACTIVITY [µmol/min/mg]
SPECIFIC ACTIVITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
additional information
-
-
The protease activity of Esp1 is required for anaphase spindle elongation in Mcd1-depleted cells (Mcd1 is a subunit of cohesin)
additional information
-
Q03018
Esp1 contributes to mitotic exit kinetics mainly through cohesin cleavage. Absence of Esp1 activity: only a minor mitotic exit delay observed which is consistent with a Cdc14 early anaphase release defect. Esp1 overexpression drives mitotic exit in Cdc20-depleted cells arrested in metaphase. Activity of overexpressed Esp1 depends on spindle integrity and the mitotic exit network.
pH OPTIMUM
pH MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
7.7
-
-
-
pH RANGE
pH RANGE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
TEMPERATURE OPTIMUM MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
37
-
-
-
TEMPERATURE RANGE
TEMPERATURE MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
30
55
-
-
SOURCE TISSUE
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
SOURCE
-
in a murine model of absent securin expression, the PTTG knock-out mouse, separase and Rad21 are over-expressed in multiple brain regions. Furthermore, Rad21 mRNA expression is highly correlated with that of securin, separase, cyclin C and sestrin 2 in fetal brains
Manually annotated by BRENDA team
-
increased expression of separase
Manually annotated by BRENDA team
-
Separase is significantly overexpressed in human breast tumors compared with matched normal tissue (Western blot)
Manually annotated by BRENDA team
-
highly expressed in human fetal cerebral cortex compared with adult
Manually annotated by BRENDA team
-
separase expression is very high in both normal nonneoplastic and neoplastic colons
Manually annotated by BRENDA team
-
separase localizes to the ingressing furrow and midbody during cytokinesis in the Caenorhabditis elegans embryo
Manually annotated by BRENDA team
-
diploid FSK3 mouse mammary epithelial cells
Manually annotated by BRENDA team
-
increased expression of separase
Manually annotated by BRENDA team
-
increased expression of separase
Manually annotated by BRENDA team
additional information
-
human separase is present in cells as a part of very large protein complex, which in addition to securin contains also Cdk and cyclin B1, both able to inhibit separase
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
localized in centrosomes before anaphase
Manually annotated by BRENDA team
-
Cut1 seems to be localized in the cytoplasm
Manually annotated by BRENDA team
-
in nondividing cells in healthy tissue, immunofluorescence microscopy
Manually annotated by BRENDA team
-
prodominant localisation
Manually annotated by BRENDA team
-
separase is abundantly expressed within the cytoplasm of a broad range of human tumor cell lines, including MDA-MB-231 breast carcinoma cells
Manually annotated by BRENDA team
-
possibly localized in the nucleus, in the mitotic stage Cut1 is mobilized to the spindle poles and the spindle microtubules
Manually annotated by BRENDA team
-
while inhibiting separase, securin is able to promote nuclear accumulation of separase
Manually annotated by BRENDA team
-
tumor cells significantly more abundant, immunofluorescence microscopy
Manually annotated by BRENDA team
additional information
-
separase localizes to cortically located filamentous structures in prometaphase I upon oocyte maturation. After fertilization, separase disappears from these structures and appears on cortical granules by anaphase I
-
Manually annotated by BRENDA team
additional information
-
Separase is present in the cell during the entire cell cycle, but excluded from the nucleus until the metaphase-anaphase transition
-
Manually annotated by BRENDA team
additional information
-
human separase is associated with centrosomes but not with spindle before anaphase, featuring predominantly cytoplasmic localization in non-dividing cells
-
Manually annotated by BRENDA team
additional information
-
budding yeast Esp1 is localized to the centrosomes and spindle before anaphase
-
Manually annotated by BRENDA team
additional information
-
in fission yeast Cut1 features similar localization in the beginning of anaphase onset persisting on the spindle until mid-anaphase
-
Manually annotated by BRENDA team
MOLECULAR WEIGHT
MOLECULAR WEIGHT MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
126000
-
-
calculated mass of the native separase, SDS-PAGE, Western blot
160000
-
-
autocleavage fragment of separase, SDS-PAGE
220000
-
-
full-length separase, SDS-PAGE
225000
-
-
recombinant separase, immunoblotting
SUBUNITS
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
?
-
x * 72900, deduced from nucleotide sequence
additional information
-
the human separase is composed of three domains: the tail, the trunk, and the head, structure modeling. The first two domains are spanned by Armadillo, ARM, repeats, which are composed of multiple 42 amino acid repeats and are present in the proteomes of all eukaryotic organisms. The ARM repeat domain is highly conserved right-handed super helix of ?-helices, which serves as molecular scaffold for protein-protein interactions. Phosphorylation and potential autocleavage sites span the region of the last ARM repeats and the central unstructured region. Human separase has an N-terminal region spanned by 26 ARM repeats and separated from the
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
proteolytic modification
F4JMQ3
during metaphase, separase is kept inactive through its binding to the chaperone securin. During anaphase, APCcdc20 cleaves securin releasing separase. Active separase cleaves itself, and the resulting N- and C-terminal fragments associate, mechanism of separase maturation during metaphase to anaphase transition
phosphoprotein
-
three phoshorylation sites within the the Cdc6-like domain of separase are identified which are critical for Cdk1 binding: Thr 1346, Thr 1363 and Ser 1399
phosphoprotein
-
in vertebrate cells, separase is phosphorylated and inhibited before anaphase by a cyclin B/CDK1. This phosphorylation leads to an inhibitory binding of the cyclin B/CDK1 complex to separase
phosphoprotein
-
phosphorylation sites span the region of the last ARM repeats and the central unstructured region
proteolytic modification
-
separase is subjected to auto-catalytic proteolytic cleavage at 3 adjacent sites, cleavage occurs specifically at anaphase coincident with separase activation, cleavage has no effect on in vitro activity
proteolytic modification
-
-
proteolytic modification
-
during metaphase, separase is kept inactive through its binding to the chaperone securin. During anaphase, APCcdc20 cleaves securin releasing separase. Active separase cleaves itself, and the resulting N- and C-terminal fragments associate, mechanism of separase maturation during metaphase to anaphase transition. Autocleavage of human separase is to be essential and results in conformational changes. The C-terminal fragment of human separase, which results from autocleavage, is more unstable than the N-terminal one. The C-terminal fragment, which possesses the catalytic domain of separase, is subjected to the N-end rule pathway of protein degradation. Consequently, catalytic activity of separase can persist only for a short period of time facilitating switch off of its proteolytic function upon entering anaphase
phosphoprotein
-
by phosphorylation of serine 1121 separase is negatively regulated
phosphoprotein
-
phosphorylation on Ser1121, mechanism to inhibit activity prior to the onset of the anaphase. Inhibitory phosphorylation of separase is not required for meiosis. Critical role of separase phosphorylation in germ cell development as well as in early embryogenesis
phosphoprotein
-
in vertebrate cells, separase is phosphorylated and inhibited before anaphase by a cyclin B/CDK1. This phosphorylation leads to an inhibitory binding of the cyclin B/CDK1 complex to separase
proteolytic modification
-
during metaphase, separase is kept inactive through its binding to the chaperone securin. During anaphase, APCcdc20 cleaves securin releasing separase. Active separase cleaves itself, and the resulting N- and C-terminal fragments associate, mechanism of separase maturation during metaphase to anaphase transition
phosphoprotein
-
most of the budding yeast cohesin is cleaved in anaphase, and this cleavage is stimulated by phosphorylation of the Scc1 subunit by the Plk1 kinase
proteolytic modification
-
during metaphase, separase is kept inactive through its binding to the chaperone securin. During anaphase, APCcdc20 cleaves securin releasing separase. Active separase cleaves itself, and the resulting N- and C-terminal fragments associate, mechanism of separase maturation during metaphase to anaphase transition
phosphoprotein
-
-
phosphoprotein
-
phosphorylation occurs under physiological conditions
proteolytic modification
-
during metaphase, separase is kept inactive through its binding to the chaperone securin. During anaphase, APCcdc20 cleaves securin releasing separase. Active separase cleaves itself, and the resulting N- and C-terminal fragments associate, mechanism of separase maturation during metaphase to anaphase transition
additional information
-
phosphorylation and potential autocleavage sites span the region of the last ARM repeats and the central unstructured region. Human separase has an N-terminal region spanned by 26 ARM repeats and separated from the two caspase-like domains, one of which is active, by the unstructured region
TEMPERATURE STABILITY
TEMPERATURE STABILITY MAXIMUM
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
4
-
-
no appreciable activity is observed at 4C
GENERAL STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
Cdc48 is required for the stability of Cut1/separase in mitotic anaphase
-
STORAGE STABILITY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
-20C, 1 week, no significant loss of activity
-
23C, 48 h, 17% loss of activity
-
4C, 48 h, 12% loss of activity
-
temperature, storage medium, duration, loss of activity
-
Purification/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
separase is expressed as a GST- or myc-tagged fusion protein in Escherichia coli BL-21 and HEK-293 cells
-
Cloned/COMMENTARY
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
depletion of endogenous separase with siRNA in NSCLCs shows that separase affects progression through the G2 phase of mitosis. Analysis of the effect of separase depletion on irradiated cells (cell cycle progression, M-phase fraction, late mitotic-linked cell death and clonogenic survival)
-
expressed in HEK-293T cells
-
expression of myc6-separase in 293T cells
-
expression of wild-type and C2029 mutant enzyme in 293 EcR and HeLa cells
-
investigation of the expression level of separase across a wide range of human tumors. Overexpression of separase transcripts strongly correlates with high incidence of relapse, metastasis and lower 5-year overall survival rate in breast and prostate cancer patients.
-
the active (S1126A/T1346A) and the inactive (C1129S) separase mutants are coexpressed in HEK-293T cells with human securin
-
analysis of correlation between overexpression of separase and aneuploidy in tetracycline-inducible diploid FSK3 mouse mammary epithelial cells
-
enhanced cyclin-dependent kinase 1 activity due to deregulation of CDC27-anaphase-promoting complex leads in turn to hyperphosphorylation of Separase, impeding chromatid separation. Regulation of mitotic progression by transforming growth factor-beta in bone marrow stromal cells is through targeting the APC-separase pathway
-
mutations introducing substitutions throughout the Esp1 polypeptide prevent loss of sister chromatid cohesion and cause mitotic failure. Generation of esp1-temperature sensitive mutant for functional analysis of further enzymes
-
Zds1 is required for Separase-induced Cdc14 activation. Analysing the role of proteins Zds1 and Zds2 in mitotic exit machinery
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generation of separase gene knock-out mutants
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continuous overexpression of cmyc tagged TbSep is lethal to the cells
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EXPRESSION
ORGANISM
UNIPROT ACCESSION NO.
LITERATURE
separase is overexpressed and mislocalized in a wide range of human cancers, including breast, prostate, and osteosarcoma
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ENGINEERING
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
C1129S
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inactive
C2029A
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no autoproteolytic cleavage, unable to cleave SCC1
C2029A
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E1483R/R1486E
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autoproteolytically cleaved like wild-type
E1503R/R1506E
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autoproteolytically cleaved like wild-type
R1486A
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mutation fails to block autocatalytical cleavage
R1506A
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mutation at autocatalytical cleavage site, mutant enzyme is still cleaved indicating the existence of a second cleavage site
R1506A/R1486A
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mutation fails to block autocatalytical cleavage
S1126A
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cleaved autoproteolytically to the same degree as wild-type, shows similar SCC1 cleavage activity as wild-type
S1126A
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a doxycycline-induced overexpression of S1126A mutant in HEK-293 cells leads to an increase of the G2/M cell population from 24 to 49% within 24 h compared to wild-type. Giemsa-stained prometaphase-chromosomes from S1126A expressing cells are mostly separated (61%), whereas those from wild-type or T1346E/T1363E/S1399D expressing cells are almost always paired (78 and 83%, respectively)
S1126A/T1346A
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active mutant
T1346A/T1363A/S1399A
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this triple mutant binds relatively more Cdk1 than S1126A but its Cdk1 binding capacity is much decreased compared to wildtype. A doxycycline-induced overexpression of the triple mutant in HEK-293 cells leads to an increase of the G2/M cell population from 24 to 37% within 24 h compared to wild-type. Giemsa-stained prometaphase-chromosomes from triple mutant expressing cells are mostly separated (61%), whereas those from wild-type or T1346E/T1363E/S1399D expressing cells are almost always paired (78 and 83%, respectively)
T1346A/T1363A/S1399D
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the mitotic arrest phenotype caused by the T1346A/T1363A/S1399A triple mutant is largely abrogated when just residue 1399 is changed to aspartate. Thus among the phosphorylation sites within the Cdc6-like domain serine 1399 is sufficient to support Cdk1-dependent inhibition of separase
T1346E/T1363E/S1399D
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changing the phosphorylation sites within the Cdc6-like domain to acidic residues results in a separase that can still be inhibited by Cdk1 in vivo
S1121A
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a mouse strain is created bearing a S1121A mutation: S1121A mutation causes infertility in mice, germ cells in the mutants are depleted during development. S1121A causes chromosome misalignment during proliferation of the postmigratory primordial germ cells, resulting in mitotic arrest, aneuploidy, and eventual cell death
S1121A
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mutation causes embryogenesis failure between the 8- and 16-cell stages in mice
S1126A
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specifically abolished separase hyperphosphorylation in Smad3-deficient cells
S1138A
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mutant containing phosphorylation site mutation compromises Cdk1-inhibitory ability
S1139A
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mutant containing phosphorylation site mutation compromises Cdk1-inhibitory ability
additional information
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RNAi of sep-1 inhibits degranulation in addition to causing extensive chromosomal segregation failures. Temperature-sensitive sep-1(e2406) allele exhibits similar inhibition of degranulation, but has minimal effects on chromosome segregation
additional information
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analysis of the homozygous zebrafish mutant cds (cease&desist), bearing a mutation in the separase gene, reveals high levels of polyploidy and aneuploidy, spindle defects, and a mitotic exit delay in mutant embryos. Carcinogenesis studies demonstrate that cds heterozygous adults have an eightfold increase in the percentage of fish bearing epithelial tumors
E1532R/R1535E
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autoproteolytically cleaved like wild-type
additional information
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in cells lacking separase or expressing noncleavable Scc1, arm cohesion is not efficiently removed during nocodazole arrest
additional information
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generation of human cells with one hESP allele-encoding uncleavable protein and another allele harboring a single cleavage site, the cells grow slowly owing to cell cycle delay, in particular during G2/M transition, but not when it was expected, i.e. during anaphase
T1346E/T1363E/S1399S
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the mitotic arrest phenotype caused by the T1346A/T1363A/S1399A triple mutant is largely abrogated when just residue 1399 is changed to serine. Thus among the phosphorylation sites within the Cdc6-like domain serine 1399 is sufficient to support Cdk1-dependent inhibition of separase
additional information
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clear evidence for a non-proteolytic function of separase is provided: by separase gene deletion in mouse oocytes it is shown that separase null-oocytes are unable to either destroy chesin along chromosome arms or segregate homologous chromosomes and these oocytes are unable to complete cell division. Microinjection of wild-type separase mRNA in separase null-oocytes restores normal homologue disjunction and polar body extrusion
H1531A
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active site point mutation prevents Scc1 from being cleaved after binding
additional information
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the loss-of-function of Esp1 activity in yeast cells could be complemented by the tobacco etch virus, TEV, protease, which is also able to cleave Scc1 thus promoting segregation of sister chromatids
additional information
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it is shown that separase acts as a direct inhibitor of cyclin-dependent kinase 1 on liberation from the inhibitory protein securin. Blocking separase-cyclin-dependent kinase 1 complex formation by microinjection of anti-separase antibodies prevents polar-body extrusion in vertebrate oocytes
APPLICATION
ORGANISM
UNIPROT ACCESSION NO.
COMMENTARY
LITERATURE
molecular biology
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separase is identified as a key cell cycle component that is required for degranulation
medicine
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the results show that separase might be a tumor supressor gene
molecular biology
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the results show that a fraction of arm cohesin is protected by Sgo1, which prevents cohesin from being removed by the prophase pathway, and that separase is partly activated in nocodazole-arrested cells and removes the arm cohesion protected by Sgo1
molecular biology
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data suggest that the co-ordinated expression of separase, securin and Rad21 is fundamental for the developing brain
molecular biology
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separse is not only required for chromosome segregation but also for meiotic exit
molecular biology
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the results indicate that inhibitory phosphorylation of separase plays a critical role in the maintenance of sister chromatid cohesion and genome stability in proliferating postmigratory primordial germ cells
molecular biology
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in Saccharomyces cerevisiae separase it is shown that separase is implicated in a second non-proteolytic pathway: separsae is essential for the activation of Cdc14 phosphatase and thus a broad programme of late mitotic events culminating in mitotic exit and cell division